Cell Line Genetics, Inc

Mycoplasma is a large group of tiny bacteria, about 0.3-0.8 um in diameter, all lacking a rigid cell wall belonging to the Mollicultes class of bacteria. Unlike other bacterial contaminates, the media will not change turbidity from a mycoplasma infection and therefore it will go undetected unless the culture is tested for mycoplasma. Due to the lack of a rigid cell wall and their small size, they are very difficult to eliminate once they contaminate a culture. Most antibiotics used in cell culturing, such as penicillin and streptomycin, work by attacking the rigid cell walls in bacteria, therefore since mycoplasma have no rigid cell wall they do not kill the mycoplasma and only mask the infection. The 0.3-0.8 um diameter and flexibility, from not having a rigid cell wall, allow these bacteria to pass through most of the conventional filters used in labs.

The two most common ways mycoplasma infection occurs:

• Contamination from an already infected cell culture
• Contamination from an infected lab member

Effects of mycoplasma on cultured cells:

• Change in growth rate and metabolism
• Change in cell morphology
• Change in the composition of the cell membrane
• Changes in gene expression
• Chromosomal aberrations
• Cell Death

How to prevent mycoplasma:

• Always use Good Cell Culture Practice (including using anti-biotic free media when antibiotics are not necessary)
• Quarantining all new cultures until mycoplasma testing has been completed
• Regularly testing for mycoplasma

Mycoplasma PCR Based Detection

Cell Line Genetics’ PCR based Mycoplasma detection is extremely sensitive, as it primes to the highly conserved 16S rRNA region of the mycoplasma’s genome. It detects 19 species of Mycoplasma and 1 species of Acholeplasma in any species of cells. Of those 19 species of mycoplasma, 6 are responsible for between 90-95% of all mycoplasma contamination in cultured stem cells.

Attributes

Extremely sensitive PCR detects only the highly conserved 16s rRNA region
Detects 19 species of Mycoplasma and 1 species of Acholeplasma
Live cells or spent media passaged twice without antibiotics accepted
Fast turnaround time, highly accurate, competitively priced

Service Specs

Used to: Detect mycoplasma contamination in cell cultures

Sample Requirements: Live cell culture (T-25 flask) or spent media (1 mL Tube) passaged twice with antibiotic free media

Species: All

Turnaround: 7-10 business days: RUSH services available
Volume/Batch Discounts are available

Cell Line Genetics offers various fluorescence in situ hybridization (FISH) services to meet your characterization needs.

Standard FISH service

Used to identify small emerging abnormal clones, complex chromosome aberrations, small partial trisomies, small marker chromosomes, and to refine chromosome breakpoints

FISH Validation of aCGH Findings

Confirm aCGH duplications and deletions using fluorescently labeled probes

Transgene Integration Site Mapping

Determine the chromosome band location of your inserted transgene

Custom FISH Probe Construction and Hybridization

Can't find the probe you are looking for? CLG can create gene specific FISH probes to meet your research need.

Array Comparative Genomic Hybridization (aCGH) is a high resolution karyotype analysis solution for the detection of unbalanced structural and numerical chromosomal alterations with high-throughput capabilities. Addition of SNPs can also detect polylpoidy, loss of heterozygosity, and uniparental disomy.

CLG offers two different array comparative genomic hybridization arrays to help characterize your cell lines. Cell Line Genetics utilizes Agilent’s Microarray platform.

SurePrint G3 Human CGH Microarray Kit, 8x60K – High-throughput, low cost alternative to karyotyping. Average coverage every 41KB with increased coverage in Refseq genes (33KB)

SurePrint G3 Human Genome CGH+SNP Microarray Kit, 4x180K – High resolution array with SNPs included. Average coverage every 25KB with increased coverage in ISCA regions (5KB)

Cell Line Genetics offers two levels of STR Profile analysis—STR100 (basic) and STR300 (comprehensive analysis). Both are designed to accommodate your needs throughout all stages of your stem cell research including cell line derivation, propagation, and long term culture. All STR services include STR profile storage in CLG's secure, encrypted database for future profile comparison analysis, and 24 hour access to reports on our secure cloud database at no additional charge. Short tandem repeat polymorphisms (STRs) are detected by utilizing Promega's PowerPlex® 16 multiplex STR system which detects STRs for 15 Codis loci plus Amelogenin—the same system used by the FBI and other government forensic labs.

Cell Line Genetics is recognized world-wide for its expertise in multispecies stem cell and cancer cell line karyotyping analysis. Unlike most cytogenetic core facilities focused on clinical testing, Cell Line Genetics is dedicated to providing the stem cell research and regenerative medicine field with the highest quality g-band karyotyping service available with fast turn-around guaranteed. Furthermore, Cell Line Genetics' scientists are rigorously trained and experienced at stem cell genomic stability analysis and identifying structural chromosomal anomalies unique to multipotent and pluripotent stem cell lines—many of which can lead to invasive cell populations. Since the company’s inception, Cell Line Genetics has characterized thousands of adult, embryonic and induced pluripotent stem cells samples from leading academic institutions, government agencies, and industry stem cell labs. Request an estimate today and have Cell Line Genetics become your g-band karyotyping quality assurance partner.

Attributes

• Harvesting of live cell cultures for chromosome analysis
• Live cell cultures and metaphase cell pellets accepted
• Chromosome analysis of 20 cells per cell line
• High quality G-band karyotypes in publication ready format
• Comprehensive interpretation of results by a CLG scientist
• Report signed by clinically certified cytogeneticist
• Results securely stored in CLG database
• 7-10 day turn around—rush services are available
• Service complies with FDA Good Laboratory Practice (GLP)

Services Specs

Used to:

• As a baseline screen:
  • when cell lines are derived
  • before the start of an experiment
  • at the conclusion of an experiment
  • before publication
  • as part of routine cell line QC
  • as part of cell banking QC
• When cultures show unusual growth properties

Detects:

• Microscopic aberrations >5Mb
• Chromosomes abnormalities:
  • aneuploidy
  • inversions
  • duplications/deletions
  • translocations

Doesn’t Detect:

• Submicroscopic aberrations <5Mb in size

Species:

• Human
• Mouse
• Various Primates
• Rat
• Cow
• Pig
• Rabbit
• Cat
• Contact us about other species

Turnaround:

• 7 - 10 business days
• RUSH services available